Identification and characterization of nuclear factor κB binding sites in the murine bcl-x promoter

J. N. Glasgow, Thomas Wood, J. R. Perez-Polo

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Signal transduction pathways that mediate neuronal commitment to apoptosis involve the nuclear factor κB (NF-κB) transcription factor. Bcl-X(L) is a potent regulator of apoptosis in the CNS and is highly expressed in the developing and adult brain. We identified three putative NF-κB DNA binding sequences clustered up-stream of the brain-specific transcription start site in the upstream promoter region. Recombinant p50/p50 and NF-κB proteins from nuclear extracts bound to these sites as determined by electrophoretic mobility shift assay and biotin-oligonucleotide/streptavidin affinity assays. NF-κB overexpression, coupled with bcl-x promoter/reporter assays using a series of murine bcl-x promoter and deletion mutants, has identified the downstream 1.1 kb of the bcl-x promoter as necessary for basal promoter activity and induction by NF-κB. The mutagenic removal of NF-κB binding sites individually or in combination revealed altered response patterns to p49/p65 and p50/p65 overexpression. These results support the hypothesis that NF-κB can act to enhance Bcl-X(L) expression via highly selective interactions, where NF-κB binding and bcl-x promoter activation are dependent on both DNA binding site sequence and NF-κB subunit composition. Our data suggest that molecular events associated with NF-κB promote regulation of neuronal apoptosis in the developing or injured CNS.

Original languageEnglish (US)
Pages (from-to)1377-1389
Number of pages13
JournalJournal of Neurochemistry
Volume75
Issue number4
DOIs
StatePublished - 2000

Fingerprint

Assays
Binding Sites
Apoptosis
Brain
Electrophoretic mobility
Signal transduction
Streptavidin
Transcription Initiation Site
DNA
Electrophoretic Mobility Shift Assay
Biotin
Nuclear Proteins
Genetic Promoter Regions
Oligonucleotides
Signal Transduction
Transcription Factors
Chemical activation
Chemical analysis
3'-(1-butylphosphoryl)adenosine

Keywords

  • Apoptosis
  • Bcl-x
  • Nuclear factor κB
  • Promoter
  • Transcription

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Identification and characterization of nuclear factor κB binding sites in the murine bcl-x promoter. / Glasgow, J. N.; Wood, Thomas; Perez-Polo, J. R.

In: Journal of Neurochemistry, Vol. 75, No. 4, 2000, p. 1377-1389.

Research output: Contribution to journalArticle

Glasgow, J. N. ; Wood, Thomas ; Perez-Polo, J. R. / Identification and characterization of nuclear factor κB binding sites in the murine bcl-x promoter. In: Journal of Neurochemistry. 2000 ; Vol. 75, No. 4. pp. 1377-1389.
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